LOCUS       sequence-458109-        7983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a version of light-inducible transcription factor EL222 
            (Val121Met) under a strong constitutive PGK1pr.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7983)
  AUTHORS   Gligorovski V, Labagnara M, Rahi SJ
  TITLE     Light-directed evolution of dynamic, multi-state, and computational 
            protein functionalities
  JOURNAL   bioRxiv 2024.02.28.582517
REFERENCE   2  (bases 1 to 7983)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.02.28.582517"
COMMENT     Sequence Label: pVG144
FEATURES             Location/Qualifiers
     source          1..7983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(13..31)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(52..68)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(52..68)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(65..87)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    76..92
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(100..106,107..124,125..130))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    145..166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(283..300)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(454..1042)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(534..553)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1213..2004,2005..2073))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1836..1855
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2074..2178)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2262..2280
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2318..2340)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2440..2459
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2815..3270)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2952..2973)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3164..3183
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3396..3418
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3410..3427
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3411..3427
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3434..3453
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3434..3452
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4860..4880
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5769..6016
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5785..5803)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(7483..7670)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
ORIGIN
        1 cagcttttgt tccctttagt gagggttaat tccgagcttg gcgtaatcat ggtcatagct
       61 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
      121 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
      181 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
      241 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
      301 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
      361 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
      421 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
      481 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      541 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      601 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      661 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      721 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      781 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      841 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      901 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      961 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1021 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1081 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     1141 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     1201 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     1261 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     1321 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     1381 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     1441 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     1501 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     1561 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     1621 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     1681 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     1741 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     1801 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     1861 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     1921 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     1981 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     2041 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     2101 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     2161 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcc tcggcgcgtg
     2221 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     2281 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     2341 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     2401 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     2461 tgagagtgca ccataccacc ttttcaattc atcatttttt ttttattctt ttttttgatt
     2521 tcggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa
     2581 ggagcacaga cttagattgg tatatatacg catattgggt cagcagtaca gaaccctcga
     2641 cgaagttgta agtggtggat cagatatcat cattgttggc agaggacttt tcgccaaggg
     2701 tagagatcct aatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac
     2761 cctatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg
     2821 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     2881 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     2941 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3001 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3061 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3121 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3181 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3241 ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg
     3301 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     3361 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     3421 ggccagtgaa ttgtaatacg actcactata gggggccagc gtggccgttt aaacatttaa
     3481 atggccggcc gctgccaccg ttgtttccac cgaagaaacc accgttgccg taaccaccac
     3541 gacggttgtt gctaaagaag ctgccaccgc cacggccacc gttgtagccg ccgttgttgt
     3601 tattgtagtt gctactgtta tttctggcac ttcttggttt tcctcttaag tgaggaggaa
     3661 cataaccatt ctcgttgttg tcgttgatgc ttaaattttg cacttgttcg ctcagttcag
     3721 ccataatatg aaatgctttt cttgttgttc ttacggaata ccacttgcca cctatcacca
     3781 caactaactt tttcccgttc ctccatctct tttatatttt ttttctggag ttcaagagaa
     3841 aaaaaaagaa aaagcaaaaa gaaaaaagga aagcgcgcct cgttcagaat gacacgtata
     3901 gaatgatgca ttaccttgtc atcttcagta tcatactgtt cgtatacata cttactgaca
     3961 ttcataggta tacatatata cacatgtata tatatcgtat gctgcagctt taaataatcg
     4021 gtgtcaagat ctgagctcca ccgcggtggc ggccgcttat ttactcagga tcccgcatgt
     4081 tgcaggtggg agttccagct cccgggaaac agctgcaaga attactcgtg agtaaggaaa
     4141 gagtgaggaa ctatcgcata cctgcattta aagatgccga tttgggcgcg aatcctttat
     4201 tttggcttca ccctcatact attatcaggg ccagaaaaag gaagtgtttc cctccttctt
     4261 gaattgatgt taccctcata aagcacgtgg cctcttatcg agaaagaaat taccgtcgct
     4321 cgtgatttgt ttgcaaaaag aacaaaactg aaaaaaccca gacacgctcg acttcctgtc
     4381 ttcctattga ttgcagcttc caatttcgtc acacaacaag gtcctagcga cggctcacag
     4441 gttttgtaac aagcaatcga aggttctgga atggcgggaa agggtttagt accacatgct
     4501 atgatgccca ctgtgatctc cagagcaaag ttcgttcgat cgtactgtta ctctctctct
     4561 ttcaaacaga attgtccgaa tcgtgtgaca acaacagcct gttctcacac actcttttct
     4621 tctaaccaag ggggtggttt agtttagtag aacctcgtga aacttacatt tacatatata
     4681 taaacttgca taaattggtc aatgcaagaa atacatattt ggtcttttct aattcgtagt
     4741 ttttcaagtt cttagatgct ttctttttct cttttttaca gatcatcaag gaagtaatta
     4801 tctacttttt acaacaaata taaaacaccg cggtggcggc cgctaaaatc atggctagcc
     4861 cgaaaaaaaa gagaaaagta gccccaccaa cggacgtttc acttggtgac gaattacatt
     4921 tagacggtga ggatgtagct atggcacatg ccgatgctct tgatgatttt gatctggaca
     4981 tgttaggtga tggggattct cccggccccg gatttacacc acatgattct gctccgtatg
     5041 gcgcgttgga catggcagat ttcgaatttg aacagatgtt tactgatgca cttggtatcg
     5101 acgaatatgg aggtgaattc ggagcagatg atactagagt tgaagtccaa cctccagccc
     5161 aatgggtatt ggatttgatc gaggcctcac ccatagcaag tgttgttagt gatcccagat
     5221 tggctgataa tcctttgatt gctatcaacc aggcctttac cgatctaact gggtattctg
     5281 aggaagaatg tgttggtcgt aattgtagat ttttggcagg atctggtact gaaccctggc
     5341 taacagataa gatccgtcaa ggtgttcgtg agcataaacc cgtgttggtt gagattttga
     5401 attacaaaaa ggatggtact ccttttcgta atgcagtttt gatggcacca atctatgatg
     5461 acgatgatga attactatac ttccttggta gccaagtgga agtggatgac gatcaaccga
     5521 acatgggcat ggcccgtaga gaacgtgctg cggagatgtt aaaaactctt tcaccacgtc
     5581 aattagaagt cacaactctg gttgcatcag gcctaagaaa taaagaagtg gctgccagac
     5641 tgggtctttc agaaaaaact gtcaaaatgc acagaggttt ggtaatggaa aaattaaact
     5701 taaaaacgag tgcagattta gttagaattg ccgtagaagc aggtatttaa gtcgacctcg
     5761 agtaatgatc atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc
     5821 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat
     5881 agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag
     5941 acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc
     6001 gaaggcttta atttgcggcc ttatcgccgg cgaataaatt aaggcgcgcc aacatatccg
     6061 gaggtaccca aagccgaatc caccacggtc accgttgttt ctgaagaaac caccacggcc
     6121 gccaccgttg ttgaaaccac cacgaggagc agaatcgcct cttctgccac caccgttacg
     6181 gatgtgaggt ggaacgtaac caccacgtct ttgttgccct tcaggcatgg aaacttgctc
     6241 tgctaactca gtcatattgg atatatgtat tactttcttt attttttgta gtttagtctt
     6301 ctatttcttt tatagtttgg tatctttttg caaaaaattt acctctatat cgtgttcgtg
     6361 cgtatttctt cctctataat atgaaaagaa aggcagtagt cttggttttc tattattcca
     6421 aatattacaa gaatacccag aaccacaaga agcagaagaa actaactgct ttatatactt
     6481 aatttcccgc cacaagctca tcgcccagaa gtttttttca gtttttcacc tgacaatctg
     6541 gcagctcgct gaaaaatttt cacctggggt cgcctgctac gtagagtgat cacgtttatg
     6601 aatagcacat tatatagtgg cccagacggt ctgatggtag ttcatgtagt taagaataaa
     6661 tactctgttt gtgtgatttc tatgtgtacg ttatctatat aaatattgtt gatggtggta
     6721 gctgtgggct gtgttctatg ctaggacacc cttagtggat ggaacgtcat cgcgacccgt
     6781 aggtgttctt gcctctctga tggcgatggc gagcgccttg aaagctgatt cggagcggtg
     6841 gtggtcgttg gtgccccgca gacagtccac atgcatggtg atacgcgcct cagtggcgaa
     6901 actctccaag aagtgcggga tcatctctgt ggatagctgg cctatgcgtt ctcgcttcag
     6961 gcccagctcc accacggcga aaggcctatt ggagaagtca accacagcgc ggctcagcgc
     7021 ctcgtccagt ggtgcaaacc catgcccgaa cctcttgata ccacgcacgg agccaagcgc
     7081 ctccttgaac gcttggccca gcgcaatccc acagtcctca acagtgtggt ggtcatcgat
     7141 gtgcaagtcc cctatacact ccaggatcag agaccacccg ccatgcttcg ccagcgcagt
     7201 caacatgtgg tccaaaaacc caactccagt gtgcacatgg atggtctgcc catcaccact
     7261 agcatattcc ttgcccagta tgctctcccg tacagaaaca gacccaccgt caagatccag
     7321 cgccagctgt atattcgtct cctgcgtaac cctcttaaca aacgccatcc tttgctcgat
     7381 gcttctcttt gcttcaatta tctcaacttt aagtaaacct gaagtaaact agaactagat
     7441 taaatcagaa ggcatccgca gctttatata catatagagc ctctttgcct tcgtttatct
     7501 tgcctgctca ttttttagta tattcttcga agaaatcaca ttactttata taatgtataa
     7561 ttcattatgt gataatgcca atcgctaaga aaaaaaaaga gtcatccgct aggtggaaaa
     7621 aaaaaaatga aaatcattac cgaggcataa aaaaatatag agtgtactag aggaggccaa
     7681 gagtaataga aaaagaaaat tgcgggaaag gactgtgtta tgacttccct gactaatgcc
     7741 gtgttcaaac gatacctggc agtgactcct agcgctcacc aagctcttaa aacgagaatt
     7801 aagaaaaagt cgtcatcttt cgataagttt ttcccacagc aaagcaatag tagaaaaaaa
     7861 caatgggaaa cgttgaatga agacaaagcg tcgtggttta aaaggaaata cgctcacgta
     7921 catgctaggg aacaggaccg tgcagcggcc agcgtggccg tttaaacatt taaatggccg
     7981 gcc
//
